Re: Live cell FRET - Glass or Plastic?

Posted by Craig Brideau on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Live-cell-FRET-Glass-or-Plastic-tp2195385p2207259.html

As everyone else is saying; glass has a much better optical quality than most plastics.  It also tends to be flatter.

Craig

On Thu, Jan 22, 2009 at 7:49 AM, Periasamy, Ammasi (ap3t) <[hidden email]> wrote:

If you are using laser as an excitation source plastic is not a good idea, you get high light scattering. If you are using arc lamp light source its ok to use good quality petri dish. It is important to understand that you are trying to collect FRET signal, as you know thats very weak signal compared to its fluorescence signal. you may see some artifacts of increase in signal in the acceptor channel when you excite with donor excitation wavelength.
Hope this helps

Ammasi Periasamy, Ph.D.
Director, Keck Center for Cellular Imaging (KCCI)
Professor of Biology and Biomedical Engineering
Biology, Gilmer Hall (064), McCormick Rd
University of Virginia
Charlottesville, VA 22904
Voice: 434-243-7602 (Office); 982-4869 (lab)
Fax:[phone]; [hidden email]
http//:www.kcci.virginia.edu
************************
Workshop on FRET Microscopy, March 3-7, 2009
http://www.kcci.virginia.edu/workshop/workshop2009/index.php
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________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Gabor Csucs [[hidden email]]
Sent: Thursday, January 22, 2009 3:35 AM

To: [hidden email]

Subject: Re: Live cell FRET - Glass or Plastic?

Hallo Pablo,

You may try the Ibidi chambers/slides although I have no idea whether
the are represented in the southern hemisphere. They have a thin plastic
bottom but concerning imaging you can reach nearly the same quality as
with glass.

Cheers    Gabor

No commercial interest.
> Hi Everyone,
>
> I'm in the process of doing FRET experiments using the pair CFP-YFP in
> live cells and I was wondering whether glass-bottomed dishes are
> better than plastic dishes for confocal imaging. HEK cells don't seem
> to want to attach to glass-bottomed dishes even when they are coated
> with poly-lysine, so using plastic dishes would make things easier.
>
> Thanks in advance,
> Pablo
>
> --
> Pablo German
>
> Plant and Food Research
> Private Bag 92169
> Auckland Mail Centre
> Auckland 1142
> New Zealand
> DDI: [phone]
> Mobile: [phone]


--
Gabor Csucs
Light Microscopy Centre, ETH Zurich
Schafmattstrasse 18, HPM F16
CH-8093, Zurich, Switzerland

Web: www.lmc.ethz.ch
Phone: [phone]
Fax: [phone]
e-mail: [hidden email]